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The quantification of PC hydroperoxide (PCOOH) in human plasma was studied by HPLC with chemiluminescence detection (HPLC-CL). We identified for the first time the monohydroperoxide of 1-palmitoyl-2-linoleoyl-PC hydroperoxide (PC 16:0/18:2-OOH) in plasma by LC-MS and HPLC-CL. The standard compound, PC 16:0/18:2-OOH (synthetic PCOOH), as well as PCOOH from egg yolk, was used. Comparison of the PCOOH concentration in each participant's plasma as determined by use of a Finepak SIL NH2 column with 2-propanol/methanol/water as the mobile phase (system A, the conventional method) gave a higher concentration than did an LC-18-DB column with methanol containing 0.01% triethylamine (system B). The mean PCOOH concentration for the 43 healthy volunteers was 55.1+/-30.4 pmol/mL (mean+/-SD) for system A and 16.3+/-9.9 pmol/mL for system B. Moreover, the main peak of the plasma extract appeared at a different time from that of synthetic PCOOH or egg yolk PCOOH in system A, whereas in system B plasma sample retention time practically corresponded to that of standard PCOOH. These findings confirm that the PCOOH plasma concentration is not so high as previously reported.

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